Abstract: A bacterium was isolated from a water sample collected from the Kephart Prong in Great Smoky Mountains National Park. This isolate did not have a conclusive match to any known bacteria in the Ribosomal Database Project (RDP); therefore, it was tested to verify it as a novel species. The full 16S rRNA gene of the newly discovered bacterium was sequenced and results showed there to be a 100% confidence that it belonged in the Enterobacteriaceae family but only an 81% confidence that is belonged in the genus Serratia suggesting that this bacterium could represent a new genus in the family. The bacterium was characterized by its preferences and tolerance to temperature, pH, and salinity as well as for metabolic capabilities such as oxidase activity, ability to hydrolyze gelatin, ability to convert tryptophan into indole, and the ability to hydrolyze casein. Antibiotic resistance was also tested against penicillin, tetracycline, chloramphenicol, clindamycin, nalidixic acid, nitrofurantoin, and colistin. Results from these phenotypic tests suggested that the isolate is not closely related to any known enterobacter species. Based on the limitations of the above methods, the full genome was sequenced to better place it in the family Enterobacteriaceae. Results of whole genome comparisons to its most closely related relatives indicate that this species is quite novel. Phylogenetics were performed and demonstrate that a new genus and species of bacteria has been cultured. This work will aid in understanding the biodiversity found in Great Smoky Mountains National Park as well as the possible contributions this bacterium provide to the ecosystem. Bacteria are also beneficial to humans and act as important resources in many different fields such as medicine, agriculture, biotechnology, and ecology. Without the knowledge and resources gained from bacteria these fields would not be as advanced as they are today. Further, conservation and restoration of ecosystems cannot occur without first understanding the biodiversity that makes up that these environments. The discovery of this isolate helps contribute to the understanding of the biodiversity in GSMNP as well as the understanding of the Enterobacteriaceae family.